Efficient Natural Plasmid Transformation ofVibrio natriegensEnables Zero-capital Molecular Biology DOI Open Access
David A. Specht, Timothy J. Sheppard, Finn Kennedy

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: Aug. 13, 2023

Abstract The fast-growing microbe Vibrio natriegens is capable of natural transformation where it draws DNA in from media via an active process under physiological conditions. Using engineered strain with a genomic copy the master competence regulator tfoX cholera combination new minimal (MCM) that uses acetate as energy source, we demonstrate naturally competent cells which are created, transformed, and recovered entirely same media, without exchange or addition media. Cells to plasmids, recombination linear DNA, co-transformation both select for scarless markerless edits. entire simple inexpensive, requiring no capital equipment room temperature (Zero Capital protocol, 10 4 cfu/ µ g), just incubator (High Efficiency 5–6 g). These retain their state when frozen transformable immediately upon thawing like typical chemical electrochemical cell. Since optimized protocol requires only 50 minutes hands-on time, V. grows quickly even on plates, started at 9 AM yields abundant culturable single colonies by 5 PM. Further, because all stages occur can be arbitrarily scaled volume, this could ideal automated directed evolution applications. As result, compete E. coli excellent chassis low-cost highly scalable synthetic biology.

Language: Английский

Synthetic biology of extremophiles: a new wave of biomanufacturing DOI
Jianwen Ye,

Yina Lin,

Xueqing Yi

et al.

Trends in biotechnology, Journal Year: 2022, Volume and Issue: 41(3), P. 342 - 357

Published: Dec. 17, 2022

Language: Английский

Citations

58
Factors affecting the competitiveness of bacterial fermentation DOI
Jong An Lee, Hyun Uk Kim, Jeong‐Geol Na

et al.

Trends in biotechnology, Journal Year: 2022, Volume and Issue: 41(6), P. 798 - 816

Published: Nov. 7, 2022

Language: Английский

Citations

32
Efficient natural plasmid transformation of Vibrio natriegens enables zero-capital molecular biology DOI Creative Commons
David A. Specht, Timothy J. Sheppard, Finn Kennedy

et al.

PNAS Nexus, Journal Year: 2024, Volume and Issue: 3(2)

Published: Feb. 1, 2024

Abstract The fast-growing microbe Vibrio natriegens is capable of natural transformation where it draws DNA in from media via an active process under physiological conditions. Using engineered strain with a genomic copy the master competence regulator tfoX cholerae combination new minimal (MCM) that uses acetate as energy source, we demonstrate naturally competent cells which are created, transformed, and recovered entirely same media, without exchange or addition fresh media. Cells to plasmids, recombination linear DNA, cotransformation both select for scarless markerless edits. entire simple inexpensive, requiring no capital equipment room temperature (zero protocol, 104 cfu/μg), just incubator (high-efficiency 105−6 cfu/μg). These retain their state when frozen transformable immediately upon thawing like typical chemical electrochemical cell. Since optimized protocol requires only 50 min hands-on time, V. grows quickly even on plates, started at 9 AM yields abundant culturable single colonies by 5 PM. Further, because all stages occur can be arbitrarily scaled volume, this could ideal automated directed evolution applications. As result, compete Escherichia coli excellent chassis low-cost highly scalable synthetic biology.

Language: Английский

Citations

7
Metabolic engineering of fast-growing Vibrio natriegens for efficient pyruvate production DOI Creative Commons
Fengli Wu, Shucai Wang, Yanfeng Peng

et al.

Microbial Cell Factories, Journal Year: 2023, Volume and Issue: 22(1)

Published: Sept. 4, 2023

Pyruvate is a widely used value-added chemical which also serves as hub of various metabolic pathways. The fastest-growing bacterium Vibrio natriegens promising chassis for synthetic biology applications with high substrate uptake rates. aim this study was to investigate if the rates V. enable pyruvate production at productivities.Two prophage gene clusters and several essential genes biosynthesis byproducts were first deleted. In order promote accumulation, key aceE encoding dehydrogenase complex E1 component down-regulated reduce carbon flux into tricarboxylic acid cycle. Afterwards, expression ppc phosphoenolpyruvate carboxylase fine-tuned balance cell growth synthesis. resulting strain PYR32 able produce 54.22 g/L from glucose within 16 h, yield 1.17 mol/mol an average productivity 3.39 g/L/h. addition, efficiently convert sucrose or gluconate titers.A novel engineered capable provide higher in This lays foundation its derivatives fast-growing natriegens.

Language: Английский

Citations

13
Ultrafast removal of toxic Cr(VI) by the marine bacterium Vibrio natriegens DOI
Xiaochen Shi, Kefan Wang, Miao Xue

et al.

Chemosphere, Journal Year: 2024, Volume and Issue: 350, P. 141177 - 141177

Published: Jan. 9, 2024

Language: Английский

Citations

4
Vibrio species as next-generation chassis for accelerated synthetic biology DOI

Changhwan Hong,

Yoojin Kim, Hyunjin Lee

et al.

Biotechnology and Bioprocess Engineering, Journal Year: 2024, Volume and Issue: 29(2), P. 241 - 253

Published: Feb. 19, 2024

Language: Английский

Citations

4
Turning waste into biofuel: Non-sterile fermentation of molasses using immobilized cells of Bacillus licheniformis BD43 for efficient 2,3-Butanediol production DOI
Mehmet Akif Omeroglu, Mustafa Özkan Baltacı, Mesut Taşkın

et al.

Industrial Crops and Products, Journal Year: 2024, Volume and Issue: 211, P. 118265 - 118265

Published: Feb. 21, 2024

Language: Английский

Citations

4
Engineered Vibrio natriegens with a Toxin–Antitoxin System for High-Productivity Biotransformation of l-Lysine to Cadaverine DOI

Shuang Zheng,

Cuihuan Zhao,

Yuemeng Chen

et al.

Journal of Agricultural and Food Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 22, 2025

Vibrio natriegens, a fast-growing bacterium, is an emerging chassis of next-generation industrial biotechnology capable thriving under open and continuous culture conditions. Cadaverine, valuable C5 platform chemical, has various chemical biological activities. This study found that V. natriegens exhibited superior tolerance to lysine, the substrate cadaverine production. For first time, synthesis pathway was introduced into for whole-cell catalysis from lysine. A high-efficiency cadaverine-producing strain harboring toxin–antitoxin system, (pSEVA341-pTac-ldcC-pHbpBC-hbpBC) with lysE (PN96_RS17440) inactivation, constructed. In 7 L bioreactors, titer increased 115 g/L in original 158 within 11 h biotransformation, exhibiting 37% increase Its productivity reached 14.4 g/L/h conversion rate as high 90%. These results confirm exceptional effective bioproduction.

Language: Английский

Citations

0
Low-biomass pyruvate production with engineered Vibrio natriegens is accompanied by parapyruvate formation DOI Creative Commons
Maurice Hädrich,

Clarissa Scheuchenegger,

Sören-Tobias Vital

et al.

Microbial Cell Factories, Journal Year: 2025, Volume and Issue: 24(1)

Published: March 28, 2025

Abstract Background Pyruvate is a precursor for various compounds in the chemical, drug, and food industries therefore an attractive target molecule microbial production processes. The fast-growing bacterium Vibrio natriegens excels with its specific substrate uptake rate as unconventional chassis industrial biotechnology. Here, we aim to exploit traits of V. pyruvate fermentations low biomass concentrations. Results We inactivated dehydrogenase complex Δ vnp12 , which harbors deletions prophage regions . resulting strain aceE was unable grow minimal medium glucose unless supplemented acetate. In shaking flasks, showed growth 1.16 ± 0.03 h − 1 produced 4.0 0.3 g Pyr L within 5 h. optimized parameters aerobic fermentation process applied constant maintenance feed 0.24 Ac resulted maximal concentration only 6.6 0.4 CDW yielded highly active resting cells (q S ) 3.5 0.2 Glc −1 41.0 1.8 volumetric productivity 4.1 Carbon balancing disclosed gap 30%, identified partly parapyruvate. Deletion ligK encoding HMG/CHA aldolase did not impact formation but plasmid-based overexpression negatively affected led 3-fold higher parapyruvate culture broth. Notably, also supernatants pyruvate-producing Corynebacterium glutamicum strain. Cell-free bioreactor experiments mimicking biological formation, pointing chemical reaction contributing synthesis. Conclusions engineered metabolically producing high at concentration. However, found that accompanied by well C. Parapyruvate seems be result conversion might supported biochemically reaction.

Language: Английский

Citations

0
β-nicotinamide mononucleotide production in Vibrio natriegens: a preliminary study DOI Creative Commons

Lan Tran,

Vu Chi Mai Tran, Huy Tuan Dat Pham

et al.

F1000Research, Journal Year: 2025, Volume and Issue: 14, P. 408 - 408

Published: April 7, 2025

Background Nicotinamide mononucleotide (NMN), is a promising nutraceutical attracting much attention for its pharmacological and anti-aging efficacies. However, NMN-containing commercial products are very high-priced due to the lack of efficient facile methods industrial-scale production. To date, various metabolic engineering strategies have been successfully applied produce NMN in Escherichia coli. Recently, Vibrio natriegens has become host bioindustry thanks rapid growth capabilities broad substrate utilization. This study aims evaluate biosynthesis capability V. natriegens. Methods Firstly, mutant strain (Δdns::araC-T7RNAP-KanR ΔpncC::FtnadE-SmR ΔnadR) was generated via multiplex genome editing by natural transformation (MuGENT). Nampt genes encoding nicotinamide phosphoribosyltransferase from Chitinophaga pinensis, Sphingopyxis sp. C-1, Haemophilus ducreyi, Vibrio phage KVP40 were codon-optimized cloned into pACYCDuet™-1 under control T7 promoter. The recombinant plasmids electroporated strain. expression NAMPTs evaluated SDS-PAGE analysis intracellular concentrations quantified HPLC. Results After two rounds MuGENT, V54-33 generated. demonstrated that all strongly expressed HPLC revealed highest concentration obtained with NAMPT pinensis (44.5 μM), followed Vibrio (23.3 μM). Conclusion feasibility natriegens.

Language: Английский

Citations

0